Lysine acetylation of aquaporin-3 promotes water reabsorption but is not essential for urine concentration

The current study aimed to decipher the role of previously discovered K282 acetylation of aquaporin-3 (AQP3) in renal water homeostasis. C57BL/6J mice harboring amino acid subsitutions to model lysine acetylation (AQP3 K282Q, termed Q mutant) or deacetylation (AQP3 K282R, termed R mutant) and wild type (WT) controls were used to determine if these mutations affected the kidney transcriptome under ad libitum pelled chow and water (standard housing condictions). Male and female mice were included in the transcriptomic analyses, and all samples were collected between 8-11 am (Zeitgeber 2-5). Wild Type (WT), acetylated mimetic (Q), and deacetylated mimetic (R), male and female mice were kept in standard housing conditions (pelleted food and tap water ad libitum; 12 h light: 12 h dark cycle). Mice were euthanized between ZT2-5 and kidneys excised, decapsulted, and snap frozen for RNA analyses.本研究旨在阐明此前发现的水通道蛋白3（aquaporin-3, AQP3）K282位点乙酰化在肾脏水稳态中的作用。本研究采用携带氨基酸替换以模拟赖氨酸乙酰化（AQP3 K282Q，命名为Q突变体）或去乙酰化（AQP3 K282R，命名为R突变体）的C57BL/6J品系小鼠，以及野生型（Wild Type, WT）对照小鼠，探究在自由采食颗粒饲料与饮水的标准饲养条件下，上述突变是否会影响肾脏转录组。转录组分析纳入雄性与雌性小鼠，所有样本均于上午8时至11时（授时因子时间2-5，Zeitgeber 2-5）采集。野生型、乙酰化模拟型（Q）与去乙酰化模拟型（R）的雌雄小鼠均饲养于标准环境：自由采食颗粒饲料与自来水，光照周期为12小时明:12小时暗。小鼠于授时因子时间2-5期间被安乐处死，摘取肾脏并剥离被膜后快速冷冻，用于RNA分析。